WSSV对锯缘青蟹的致病性及血清酶指标影响
STUDIES ON PATHOGENICITY OF WHITE SPOT SYNDROM VIRUS AND EFFECT ON HEMOLYMPH ENZYMES ACTIVITIES CHANGES IN MUD CRAB SCYLLA SERRATA (FORSKAL)
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摘要: 锯缘青蟹(Scylla serrata)俗称青蟹,是我国重要的海水养殖蟹类。近年来,浙江、福建、广东等青蟹主要养殖地区出现了严重的青蟹病害。对浙江省养殖青蟹的发病原因和流行病学调查发现,白斑综合征病毒(White spotsyndrome virus,WSSV)与青蟹发病存在较大相关性。为进一步研究WSSV对青蟹的致病性和发病机理,作者采用白斑综合征病毒的除菌过滤液,以1∶10—1∶10000稀释度注射感染青蟹,结果表明1∶10、1∶100感染组的青蟹死亡率达100%,1∶1000感染组死亡率为66.7%,1∶10000感染组死亡率为38.9%。根据攻毒悬液的病毒浓度计算出WSSV对青蟹的LD50为1.19×106拷贝/只(7.93×103拷贝/g组织);取WSSV感染青蟹血淋巴进行PCR检测,攻毒死亡青蟹的WSSV检出率为100%,表明WSSV对青蟹有很强的致病力。分析病毒感染濒死蟹的血清酚氧化酶(PO)、过氧化物酶(POD)、超氧化物歧化酶(SOD)、碱性磷酸酶(ALP)、谷丙转氨酶(GPT)、谷草转氨酶(GOT)等主要酶指标,发现病毒感染青蟹的PO、POD和SOD活力明显低于对照组,而ALP、GPT和GOT的活力则明显高于对照组;用WSSV单克隆抗体对感染蟹进行免疫组化分析,发现WSSV主要侵染青蟹的鳃、甲壳下表皮、心脏、肠、胃等组织的上皮细胞,尤其以鳃上皮细胞损害最为严重。Abstract: Mud crab,Scylla serrata(Forskal),is main economically cultivated marine crab in China,with production of 108,500 tons,88.14%of the total production of the world.The main mud crab region distributed along the coastline of Southeast China,including Zhejiang,Fujian,Guangdong,Guangxi and Hainan Province.Recently,serious diseases occurred in main mud crab culture region,with the morbidity of 13.77% in cultivated mud crab in 2006 in whole China,resulting in 32.71 million dollars lost.White spot syndrome virus(WSSV),the most pathogen of shrimp culture in China,was considered an important pathogen in cultivated mud crab.It showed that WSSV detection with PCR was closely relative with occurring of mud crabs diseases during the epidemics investigations from 2006 to 2007.To further study the pathogenicity of WSSV to mud crab,WSSV bacteria-free suspension,collected from typical WSSV infected black tiger shrimp(Penaeus monodon),was used for challenging to healthy mud crab by intramuscular injection at 10-folded dilutions ranged from 1∶10 to 1∶10000.The results showed that the cumulative mortalities for dilutions of 1∶10 to 1∶100,1∶1000,1∶10000 were 100%,66.7% and 38.9% respectively.The LD50 of WSSV to mud crab was calculated as 1.19×106 copies/crab,or 7.93×103 copies/g of crab weight by Muench-Reed methods.Gills and hemolymph of moribund mud crabs challenged with WSSV were positive by PCR.These results indicated that WSSV was highly virulent to mud crab.The activities of main hemolymph enzymes,including phenoloxidase(PO),peroxidase(POD),superoxide dismutase(SOD),alkaline phosphatase(ALP),glutamate-pyruvate transaminase(GPT) and glutamic-oxaloacetic transaminase(GOT) of mud crab were analyzed.WSSV infected moribund crabs showed significantly descent in activities of PO,POD and SOD,while ascent in ALP,GPT and GOT compared with the control group.WSSV mainly distributed in epithelia of gills,cuticular of skeletal plate,heart,intestine and stomach by immunohistochemistry with Mabs against WSSV,and gill epithelia showed heavily damaged with the main distribution of WSSV in nucleic.It can be concluded that WSSV was a main pathogen of mud crab with high pathogenicity.
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Keywords:
- Scylla serrata /
- WSSV /
- Pathogenicity /
- Enzyme activity
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